Abstract
Targeting altered cancer cell metabolism with the glycolysis inhibitor, 2-deoxyglucose (2DG), is a viable therapeutic strategy, but the effects of 2DG on lymphoma cells and the mechanism of action are unknown. Five T-cell lymphoma lines and two B-cell lymphoma lines were shown to be highly sensitive to 2DG. Examination of the cell death pathway demonstrated pro-apoptotic protein Bax ‘activation’ and caspase cleavage in 2DG-treated cells. However, Q-VD-OPh, a potent inhibitor of caspase activity provided minimal protection from death. In contrast, overexpressing the anti-apoptotic protein Bcl-2 dramatically enhanced the survival of 2DG-treated cells that was negated by a Bcl-2 antagonist. BH3-only members, Bim and Bmf, were upregulated by 2DG, and shRNAs targeting Bim protected from 2DG toxicity demonstrating that Bim is a critical mediator of 2DG toxicity. 2DG also induced GADD153/CHOP expression, a marker of endoplasmic reticulum (ER) stress and a known activator of Bim. Mannose, a reagent known to alleviate ER stress, transiently protected from 2DG-induced cell death. Examination of the effects of 2DG on energy metabolism showed a drop in ATP levels by 30 min that was not affected by either Bcl-2 or mannose. These results demonstrate that ER stress appears to be rate limiting in 2DG-induced cell death in lymphoma cells, and this cell killing is regulated by the Bcl-2 family of proteins. Bcl-2 inhibition combined with 2DG may be an effective therapeutic strategy for lymphoma.
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Acknowledgements
This work was supported by RO1 no. CA104695, RO1 no. CA133114, R01 no. DK084058, and R01 no. GM086389. We thank Dr Chris van de Wetering for assistance in generating lymphoma cell lines, Dr Siegfried Janz and Dr SS Han for B-cell neoplasms, Dr Agshin Taghiyev and Dr Van Tompkins for their advice and assistance with TCL transduction experiments, Dr Craig Kuder and Dr Ray Hohl for antibodies and assistance with ER-stress markers, Heather Tyra for assistance with RT–PCR, Han Du for advice with the Bcl-2 IP experiments, Rebecca Glover and Jacob Wolf for their technical advice on western blotting and Dr Craig Thompson (The Sloan-Kettering Cancer Center) for Bax/Bak double-knockout cells.
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Zagorodna, O., Martin, S., Rutkowski, D. et al. 2-Deoxyglucose-induced toxicity is regulated by Bcl-2 family members and is enhanced by antagonizing Bcl-2 in lymphoma cell lines. Oncogene 31, 2738–2749 (2012). https://doi.org/10.1038/onc.2011.454
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DOI: https://doi.org/10.1038/onc.2011.454
